An image analysis strategy to determine the distribution of cell types in spleen sections

An image analysis strategy was designed to objectively determine distributi on patterns of cell types in spleen sections. The strategy was applied to r at spleen cryostat sections that were stained immunohistochemically by mean s of monoclonal antibodies against different populations of lymphocytes and macrophages. The strategy revealed three segments of the spleen beginning in the middle of a central arteriole and ending within the red pulp. In eac h of these segments, three consecutive zones were established: the white pu lp, the marginal zone, and the red pulp. In each tissue section, three segm ents were selected starting in two different arterioles. Consequently, six segments were analysed in each section. Special software was used to calcul ate percentages of positive staining in all zones in each segment. Image an alysis data for each monoclonal antibody tested correlated closely with mic roscopical observations. The proposed strategy allows objective quantificat ion of lymphocyte and macrophage populations and their distribution pattern s. It is an useful tool for studying imbalances in cell populations in the spleen due to immune challenges.